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Related Experiment Video

Updated: Jun 23, 2026

Selective Labelling of Cell-surface Proteins using CyDye DIGE Fluor Minimal Dyes

Selective Labelling of Cell-surface Proteins using CyDye DIGE Fluor Minimal Dyes

Published on: November 26, 2008

A fluorescent codetection system for immunoblotting and proteomics through ECL-Plex and CyDye labeling.

Ciara A McManus¹, Pamela M Donoghue, Michael J Dunn

¹Proteome Research Centre, UCD Conway Institute of Biomolecular and Biomedical Research, Dublin 4, Ireland. ciara.mcmanus@ucd.ie

Methods in Molecular Biology (Clifton, N.J.)

|April 21, 2009

Summary

Differential gel electrophoresis (DIGE) combined with ECL-Plex CyDye antibodies advances western blotting. This technique allows simultaneous visualization of total protein profiles and specific protein detection for improved validation.

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Area of Science:

Proteomics
Biochemistry
Molecular Biology

Background:

Differential gel electrophoresis (DIGE) has revolutionized proteome analysis with its sensitive CyDye labeling.
Conventional western blotting faces limitations in simultaneous protein profiling and specific detection.

Purpose of the Study:

To advance western blotting by combining DIGE labeling with ECL-Plex CyDye conjugated secondary antibodies.
To enable simultaneous visualization of total protein expression and specific immunodetection for protein validation.

Main Methods:

Utilized 1-D and 2-D electrophoresis.
Employed DIGE labeling combined with ECL-Plex CyDye conjugated secondary antibodies.
Detected heat shock protein 27 (hsp 27) in a human cardiac protein extract.

Main Results:

Demonstrated a simple, rapid, and reproducible technique for protein detection.
Successfully visualized total protein expression alongside specific immunodetection of hsp 27.
Confirmed the location of hsp 27 within a complex cardiac protein sample.

Conclusions:

The combined DIGE and ECL-Plex CyDye method significantly aids protein validation after 2-D gel separation.
This technique offers simultaneous total protein profiling and specific protein immunodetection.
The method is effective for analyzing specific proteins, such as hsp 27, in complex biological samples.